
瓦莱氏芽孢杆菌YH-18发酵液的喷雾干燥工艺
Spray drying processes of Bacillus valeriana YH-18
【目的】瓦莱氏芽孢杆菌(Bacillus valeriana) YH-18菌株是1株对多种林木具有抗病促生效果的优良菌株。菌粉是生物菌剂应用在生产上的一种类型,以高集粉率和活菌量作为主要目标,确定YH-18菌粉制备的最佳喷雾干燥的工艺条件,为抗病促生瓦莱氏芽孢杆菌YH-18的规模化应用提供技术支撑。【方法】利用喷雾干燥法对YH-18菌株的发酵液进行喷雾干燥,采用单因素试验和正交试验设计,研究了瓦莱氏芽孢杆菌YH-18菌粉制备的喷雾干燥工艺条件。【结果】瓦莱氏芽孢杆菌YH-18发酵液喷雾干燥的最佳工艺条件为:进风口温度95 ℃,最佳保护剂为15%脱脂奶粉,雾化压力0. 25 MPa,进样速度10 mL/min,真空度-0. 03 Pa。在该条件下喷雾干燥得到的YH-18固体菌剂产物,活菌量可以达到3. 28×10 9 cfu/g。YH-18固体菌剂对林木溃疡病病原菌拟茎点霉(Phomopsis sp.)保持有明显拮抗作用。【结论】固体菌剂相对于液体菌剂具有更易储藏、便于运输、货架期长等优点。喷雾干燥法具有控制性强、稳定性高的特点,是实现液体菌剂到固体菌剂的有效途径。YH-18发酵液喷雾干燥工艺条件的确定为未来规模化固体菌剂应用提供了技术支持。
【Objective】Bacillus valeriana YH-18 is a bacterial strain that exhibits an excellent antifungal ability in different kinds of trees. This strain is used as a biological agent in the form of bacterial powder. In this study, the spray drying of YH-18 strain was investigated as a method for producing the bacterial powder.【Method】The conditions of spray drying of YH-18 strain were studied using single-factor and orthogonal tests that were designed to attain a high powder collection rate and number of living bacteria as the outcomes.【Result】The best conditions for the spray drying of B. valeriana YH-18 fermentation liquid are as follows: air inlet temperature, 95 ℃; addition of 15 % skim milk powder; atomization pressure, 0.25 MPa; feeding speed, 10 mL/min; and vacuum degree, -0.03 Pa. Under these conditions, the yield of YH-18-based solid product obtained after the spray drying of B. valeriana YH-18 fermentation liquid reached up to 3.28 × 10 9 cfu/g. The obtained solid fungicide provided potent protection against the disease caused by Phomopsis sp., a pathogen of wood canker disease.【Conclusion】Solid fungicides possess several advantages over liquid fungicides, such as ease of storage and transportation and long shelf life. The spray drying method can be applied in a highly controlled manner and has been shown to impart increased stability to the end product. It is an effective way of converting liquid fungicides to solid fungicides. Therefore, the determination of spray drying conditions of B. valeriana YH-18 fermentation liquid will provide further technical support in the application and production of large-scale solid fungicides in future.
瓦莱氏芽孢杆菌 / 固体菌剂 / 喷雾干燥 / 工艺条件 / 活菌量 / 集粉率
Bacillus valeriana / microbial solid fungicides / spray drying / process parameters / viable cell counts / collector efficiency
[1] |
吴建峰, 林先贵 . 我国微生物肥料研究现状及发展趋势[J]. 土壤, 2002,34(2):68-73.
|
[2] |
Bacillus methanolicus can efficiently utilize methanol as a sole carbon source and has an optimum growth temperature of 50 degrees C. With the exception of mannitol, no sugars have been reported to support rapid growth of this organism, which is classified as a restrictive methylotroph. Here we describe the DNA sequence and characterization of a 19,167-bp circular plasmid, designated pBM19, isolated from B. methanolicus MGA3. Sequence analysis of pBM19 demonstrated the presence of the methanol dehydrogenase gene, mdh, which is crucial for methanol consumption in this bacterium. In addition, five genes (pfk, encoding phosphofructokinase; rpe, encoding ribulose-5-phosphate 3-epimerase; tkt, encoding transketolase; glpX, encoding fructose-1,6-bisphosphatase; and fba, encoding fructose-1,6-bisphosphate aldolase) with deduced roles in methanol assimilation via the ribulose monophosphate pathway are encoded by pBM19. A shuttle vector, pTB1.9, harboring the pBM19 minimal replicon (repB and ori) was constructed and used to transform MGA3. Analysis of the resulting recombinant strain demonstrated that it was cured of pBM19 and was not able to grow on methanol. A pTB1.9 derivative harboring the complete mdh gene could not restore growth on methanol when it was introduced into the pBM19-cured strain, suggesting that additional pBM19 genes are required for consumption of this carbon source. Screening of 13 thermotolerant B. methanolicus wild-type strains showed that they all harbor plasmids similar to pBM19, and this is the first report describing plasmid-linked methylotrophy in any microorganism. Our findings should have an effect on future genetic manipulations of this organism, and they contribute to a new understanding of the biology of methylotrophs.
|
[3] |
尹向田, 杨阳 . 甲基营养型芽孢杆菌GSBM05产抗菌活性物质发酵条件优化[J]. 江苏农业科学, 2018,46(20):89-93.
|
[4] |
赵旭, 常思静, 景春娥 , 等. 我国植物内生菌研究进展[J]. 中国沙漠, 2010,30(1):87-91.
|
[5] |
张妙宜, 云天艳, 周登博 , 等. 甲基营养型芽孢杆菌的研究进展[J]. 热带农业科学, 2017,37(9):66-71.
|
[6] |
江明明 . 几株优良内生菌的分离及鉴定[D]. 南京:南京林业大学, 2016.
|
[7] |
魏奇 . 几株优良内生细菌在上海生态林上的应用研究[D]. 南京:南京林业大学, 2017.
|
[8] |
徐鹏, 王利莹, 曹小勇 , 等. 乳化喷雾干燥法制备戊唑醇/壳聚糖微胶囊[J]. 南京林业大学学报(自然科学版), 2017,41(3):157-161.
|
[9] |
贺娜, 于晓晨, 于才渊 . 喷雾干燥技术的应用[J]. 干燥技术与设备, 2009,7(3):116-119.
|
[10] |
黄敏, 姚莉, 赵玲华 . 喷雾干燥法制备胡麻籽油微胶囊工艺的优化[J]. 农产品加工, 2015(7):42-45.
|
[11] |
孟祥坤, 于俊杰, 尹小乐 , 等. 拮抗细菌枯草芽孢杆菌T429喷雾干燥工艺研究[J]. 中国生物防治学报, 2014,30(1):101-106.
本研究前期成功分离获得1株对水稻真菌病害具有较强抑制作用的枯草芽孢杆菌T429,为开发生防菌农药新剂型,利用喷雾干燥法对生防菌T429发酵液进行处理,以喷雾干燥后产物活菌量为主要指标,采用单因素和正交试验法,对影响喷雾干燥产物指标的主要因素进行优化。结果表明,最佳喷雾干燥条件为进风温度120℃、进样速度720 mL/h、雾化口径1 mm、雾化压力0.1 MPa、热风流量30 M3/h,可获得产物活菌量达8.61×109 cfu/g。该工艺稳定、可控,为以枯草芽孢杆菌为核心的微生物杀菌剂新剂型的研发提供了科学依据。
|
[12] |
周竞, 程辉彩, 崔冠慧 , 等. 枯草芽孢杆菌BSD-2菌株喷雾干燥工艺优化[J]. 现代农药, 2016,15(1):13-15,27.
|
[13] |
区伟佳, 王荣, 李华兴 , 等. 高浓度大豆生防芽孢杆菌粉剂喷雾干燥微胶囊化工艺研究[J]. 大豆科技, 2010(3):3-6.
确定了微胶囊材料麦芽糊精和阿拉伯胶与大豆生防芽孢杆菌AF67具有良好的生物相容性.应用正交试验确定了生防菌喷雾干燥过程中影响菌粉收率、存活率、含水量的主要因素,并建立多元指数模型.模型确定的最佳工艺条件为:进风温度170℃,进料速度10ml/min,芯材与填充荆配比为1:8,固型物含量25%,活菌数达2.1×1010cfu/g.经验证,该数学模型具有可靠性.
|
[14] |
刘庆莉, 王金生, 刘丽君 , 等. 大豆根瘤菌剂载体的选择及最佳施用浓度筛选[J]. 大豆科学, 2014,33(2):207-210.
|
[15] |
赵斌, 张建志, 李妍 , 等. 保加利亚乳杆菌冷冻干燥保护剂的研究[J]. 中国乳品工业, 2013,41(2):10-12.
|
[16] |
郑唯, 朱丹, 牛广财 , 等. 富糖类物料喷雾干燥热熔性粘壁现象研究进展[J]. 农业科技与装备, 2018(1):57-58,61.
|
[17] |
李春梅, 迟玉杰, 王丹 , 等. 改性甜玉米浆喷雾干燥条件的探讨[J]. 食品科技, 2012,37(3):192-196.
|
[18] |
刘军辉, 李利 . 我国微生物肥的应用研究进展[J]. 河北果树, 2018(5):5-6.
|
[19] |
高书锋, 刘惠知, 胡新旭 , 等. 畜禽用凝结芽孢杆菌菌剂制备工艺的初步研究[J]. 饲料工业, 2012(13):37-40.
|
/
〈 |
|
〉 |